BvM14-RIN4 Gene of Sugar Beet M14 Line: Cloning and Response to Salt Stress

doi:10.11924/j.issn.1000-6850.casb2020-0848

Abstract

Abstract:

To study the salt-resistant function of RIN4 gene in sugar beet monosomic additional line M14 line, sugar beet M14 line was used as experimental material, the full-length cDNA sequence of the BvM14-RIN4 gene was obtained by PCR, and the BvM14-RIN4 gene was analyzed by bioinformatics and its tissue specificity and response to salt stress were explored. The results showed that the ORF length of BvM14-RIN4 gene was 264 bp which encoded 87 amino acids, the molecular weight of the protein was 9.67 kDa, the theoretical pI was 9.64, and the protein was preliminarily identified as a hydrophilic protein. The secondary structure of BvM14-RIN4 protein was mainly composed of extension chain and random curl; the tertiary structure of BvM14-RIN4 protein predicted the conserved domain and hydrophobic region of the protein; the phylogenetic tree showed that BvM14-RIN4 protein was closely related to RIN4 protein of Spinacia oleracea L. and Chenopodium quinoa L.. The results of quantitative RT-PCR showed that the expression level of BvM14-RIN4 gene in roots leaves was 2.26 times that of leaves, which indicated that it had tissue specificity; the expression of BvM14-RIN4 gene was up-regulated in roots under 200 mmol/L NaCl, which indicated that it was involved the response of sugar beet M14 line to salt stress. This study is of certain significance in mining high quality genes of sugar beet M14 line and carrying out genetic improvement of sugar beet, the results could provide important references for studying the function of BvM14-RIN4 gene.

Key words: sugar beet M14 line, BvM14-RIN4, gene clone, bioinformatics analysis, quantitative RT-PCR

CLC Number:

S566.3

Jia Huixu, Li Haiying, Duanmu Huizi. BvM14-RIN4 Gene of Sugar Beet M14 Line: Cloning and Response to Salt Stress[J]. Chinese Agricultural Science Bulletin, 2021, 37(14): 27-33.

Figures/Tables 10

References 23

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引物名称	序列
BvM14-RIN4-S	5'-TGTTTCTTGTATTACTTTTGAGAGC-3'
BvM14-RIN4-AS	5'-TCAAAGCCACCTTCTACTACAAAT-3'
qRT-PCR-RIN4-S	5'-GCAGGTAACAGATCCTTGCCA-3'
qRT-PCR-RIN4-AS	5'-TTAGGGCTCTGGTTAACGCC-3'

引物名称	序列
BvM14-RIN4-S	5'-TGTTTCTTGTATTACTTTTGAGAGC-3'
BvM14-RIN4-AS	5'-TCAAAGCCACCTTCTACTACAAAT-3'
qRT-PCR-RIN4-S	5'-GCAGGTAACAGATCCTTGCCA-3'
qRT-PCR-RIN4-AS	5'-TTAGGGCTCTGGTTAACGCC-3'